Evaluating the reliability of microsatellite genotyping from low-quality DNA templates with a polynomial distribution model  

作　　者：HE Gangi HUANG Kang GUO SongTao JI WeiHong QI XiaoGuang REN Yi JIN XueLin LI BaoGuo 

机构地区：[1]Key Laboratory of Resource Biology and Biotechnology in Western China of Ministry of Education, College of Life Sciences, Northwest Univer-sity, Xi'an 710069, China [2]Ecology & Conservation Group, Institute for Natural Sciences, Massey University, Auckland 904, New Zealand [3]Institute of Zoology, Shaanxi Academy of Sciences, Xi'an 710032, China [4]Shaanxi Wild Animal Rescue and Research Center, Xi'an 710402, China

出　　处：《Chinese Science Bulletin》2011年第24期2523-2530,共8页

基　　金：supported by the National Natural Science Foundation of China (30970379, 30970444 and 30970168);the Natural Science Foundation of Shaanxi Province (2009JQ3001);the Scientific Research Foundation of the Educa-tion Department of Shaanxi Province (09JK748);the Opening Foun-dation of the Key Laboratory of Resource Biology and Biotechnology in Western China (Northwest University) of Ministry of Education

摘　　要：Molecular studies using trace DNA, such as from museum specimens, ancient or forensic samples and samples obtained noninvasively, often have a common problem of low quality of DNA templates. Amplification errors, such as allelic dropout and false allele, may arise during polymerase chain reaction (PCR) using such samples. A mathematical model which treats homozygotes and heterozygotes discriminately has been developed to measure sample quality and compute the confidence level of using multipletube approaches. We use plucked hair samples collected from 26 individual Sichuan snub-nosed monkeys (Rhinopithecus roxel- lana) to test the model. In this case, a confidence level of 99% can be achieved by three positive PCRs. If the sample quality is very poor and requires many PCR replicates, an alternative multiplestep genotyping method is recommended. This model enables researchers to optimize experimental protocols through pilot studies and obtain reliable genetic information using noninvasive sampling method.Molecular studies using trace DNA, such as from museum specimens, ancient or forensic samples and samples obtained noninva- sively, often have a common problem of low quality of DNA templates. Amplification errors, such as allelic dropout and false allele, may arise during polymerase chain reaction （PCR） using such samples. A mathematical model which treats homozygotes and heterozygotes discdminately has been developed to measure sample quality and compute the confidence level of using multiple- tube approaches. We use plucked hair samples collected from 26 individual Sichuan snub-nosed monkeys （Rhinopithecus roxel- lana） to test the model. In this case, a confidence level of 99% can be achieved by three positive PCRs. If the sample quality is very poor and requires many PCR replicates, an alternative multiple-step genotyping method is recommended. This model enables researchers to optimize experimental protocols through pilot studies and obtain reliable genetic information using noninvasive sampling method.

关 键 词：DNA模板 基因分型 低质量 可靠性评估 分布模型 微卫星 多项式 PCR产物 

分 类 号：Q75[生物学—分子生物学]